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甘菊cDNA-AFLP反应体系的优化
引用本文:黄河,王顺利,曹华雯,戴思兰.甘菊cDNA-AFLP反应体系的优化[J].生物技术通报,2009(11).
作者姓名:黄河  王顺利  曹华雯  戴思兰
作者单位:1. 北京林业大学园林学院,北京,100083
2. 北京林业大学园林学院,北京100083;北京农学院城乡发展学院,北京,102206
基金项目:国家高技术研究发展计划("863"计划)重点项目 
摘    要:以甘菊为试验材料,研究了影响cDNA-AFLP反应体系的几个关键因素,建立了适宜甘菊的cDNA-AFLP分析体系,并得到了清晰可辨的cDNA-AFLP指纹图谱.结果表明:适用于甘菊叶片总RNA提取的方法为改进的Trizol法;酶切连接采用一步法,dscDNA酶切用量为300 ng,酶切连接时间为8 h;PCR选择性扩增反应时,反应体系中最佳组合为:引物浓度0.4 mM、Mg2+浓度1.25 mM、Taq酶浓度0.9 U、dNTP浓度0.3 mM.

关 键 词:甘菊  反应体系优化

Optimization of cDNA-AFLP Technical System for Chrysanthemum lavandulifolium
Huang He,Wang Shunli,Cao Huawen,Dai Silan.Optimization of cDNA-AFLP Technical System for Chrysanthemum lavandulifolium[J].Biotechnology Bulletin,2009(11).
Authors:Huang He  Wang Shunli  Cao Huawen  Dai Silan
Abstract:After systematical studies on the factors affecting cDNA-AFLP analysis system of Chrysanthemum lavandulifolium,an optimized system was established which showed a distinct electrophoretogram of cDNA-AFLP. The results indicated that using the improved Trizol method can isolate RNA quickly and efficiently from Chrysanthemum lavandulifolium; cDNA was digested and ligated by one step. The dosage of dscDNA was 300 ng. Digestion and ligation was performed in 8 hours; in 20 ul PCR reaction system of selective amplification, the primer concentration was 0. 4 mM; dNTP concentration was 0. 3 mM; Mg~(2+) concentration was 1. 25 mM; Taq DNA concentration was 0.9 U.
Keywords:cDNA-AFLP
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